Anisotropia de fluorescência da sonda AHBA em lipossomas de misturas lipídicas e contendo colesterol
DOI:
https://doi.org/10.29384/rbfm.2024.v18.19849001755Palavras-chave:
fluorescência, biofísica, lipossomas, misturas lipídicas, colesterolResumo
Sondas fluorescentes hidrofóbicas no estudo de membranas modelo têm sido amplamente utilizadas, pois apresentam dinâmicas rotacionais diferentes para diversos ambientes lipídicos, sendo capazes de monitorar a fluidez da bicamada lipídica e os mecanismos dependentes deste fator. Entretanto, a eficácia do uso de sondas fluorescentes anfipáticas deve ser avaliada uma vez que a localização dessas sondas na membrana pode causar uma falha no monitoramento do empacotamento das cadeias graxas. Foram apresentados, neste trabalho, resultados da fluorescência estática da sonda AHBA (2-Amino-N-hexadecilbenzamida) utilizada em estudos do comportamento de fase de vesículas lipídicas (lipossomas) formadas por misturas lipídicas e contendo colesterol. Foram variados, na produção dos lipossomas, o comprimento da cadeia hidrocarbônica dos lipídeos, bem como o tipo de cabeça polar: fosfatidilcolina (PC) e fosfatidilglicerol (PG). Parâmetros como intensidade de fluorescência, deslocamento espectral e anisotropia estática foram obtidos para o AHBA, inserido nas diferentes bicamadas lipídicas. A anisotropia estática se mostrou o parâmetro mais adequado no monitoramento da transição da fase gel para a fase fluida da bicamada, detectando ainda a presença de colesterol no sistema, enquanto os resultados de deslocamento espectral mostraram que o AHBA possui baixa sensibilidade a alterações de polaridade discretas no ambiente ao seu redor.
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